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Helicobacter spp. infections in mice can have broad-ranging effects on gastrointestinal, reproductive, and immune systems. This can introduce significant confounding variables for research and may reduce scientific rigor. Screening mouse colonies for Helicobacter species can be accomplished via noninvasive PCR testing on filter paper placed in animal-free dirty bedding sentinel cages. In our facility, one tablespoon of dirty bedding from each cage on a rack is added to a designated sentinel cage every 3 wk at cage change, and PCR testing is performed on in-cage filter paper quarterly. We hypothesized that cages that received Helicobacter spp.-positive bedding at later time points would have a lower detection rate of Helicobacter spp. with PCR testing compared with cages that received positive bedding at earlier time points due to the filter paper becoming saturated. To determine if screening would be able to detect one positive row of cages on a rack, 9 tablespoons of Helicobacter- positive bedding and 71 tablespoons of negative bedding were added at the 3-, 6-, or 9-wk time points to 14 empty sentinel cages per time point. Negative bedding was added every 3 wk to cages not scheduled to receive positive bedding. Negative controls received 80 tablespoons of negative bedding and positive controls received 80 tablespoons of positive bedding at each time point. Filter paper was tested via PCR for Helicobacter spp. at 12 wk. All positive controls tested positive, and all negative controls tested negative. Two 3-wk cages, two 6-wk cages, and three 9-wk cages were positive, indicating no difference between time points. This resulted in a 16.7% Helicobacter spp. detection rate. These results indicate that PCR on in-cage filter paper may not be reliable in detecting low levels of Helicobacter spp. nucleic acid in dirty bedding.

Gastric ulcer is a common disease affecting pigs worldwide, with a prevalence reported as high as 93%. The cause of porcine gastric ulcer is multifactorial, with Helicobacter suis (H. suis) being considered as the primary pathogenic factor. To date, prevalence of H. suis resulting in porcine gastric ulcer in Taiwan has not been investigated. In this study, we collected 360 pig stomachs from the slaughterhouses. In addition, stomach tissues from the 88 diseased pigs submitted for necropsy were divided into symptomatic and asymptomatic groups. Gastric lesions were scored, and polymerase chain reaction was used to determine the occurrence of gastric ulcer and the prevalence of H. suis. The positive rate of H. suis in the samples from slaughtered pigs was 49.7%, and both infection of H. suis and the presence of gastric lesions were prone to occur in autumn. The positive rates of H. suis infection in the symptomatic and asymptomatic groups were 59.1% and 31.8%, respectively. Moreover, the proportion of the samples with gastroesophageal ulcer in the symptomatic group was 68.2%, predominantly observed in growing pigs. The incidence of the samples from the slaughterhouses with gastroesophageal erosion to ulceration revealed a significant difference between H. suis -infected and H. suis -uninfected pigs; however, there is no significant difference in the samples of diseased pigs. In conclusion, H. suis infection was associated with gastric ulcer in slaughtered pigs, but it was not the primary cause of gastroesophageal ulcer in diseased pigs with clinical symptoms.

Helicobacter species (spp.) is a gram-negative spiral-shaped motile bacterium that causes gastritis in pigs and also colonizes in the human stomach. The present study assessed the prevalence of Helicobacter spp. in pig gastric mucosa and the stool of pig farmers in Assam, India. A total of 403 stomach samples from pig slaughter points, 74 necropsy samples of pigs from pig farms, and 97 stool samples from pig farmers were collected. Among the pig stomach samples, 43 (20.09%) of those with gastritis showed the presence of Gram-negative, spiral-shaped organisms, while only 3.04% of stomach samples without lesions had these organisms. Scanning Electron Microscopy (SEM) of urease-positive stomach samples revealed tightly coiled Helicobacter bacteria in the mucus lining. Histopathological examination showed chronic gastritis with hemorrhagic necrosis, leucocytic infiltration, and lymphoid aggregates. PCR confirmed the presence of Helicobacter suis in 19.63% of pig stomach samples and 2.08% of pig farmer stool samples. Additionally, 3.12% of the stool samples from pig farmers were positive for Helicobacter pylori. Phylogenetic analysis revealed distinct clusters of Helicobacter suis with other Helicobacter spp. These findings highlight the prevalence of Helicobacter in both pig gastric mucosa and pig farmer stool. The findings highlight the need for improved sanitation and hygiene practices among pig farmers to minimize the risk of Helicobacter infection in humans., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigated the role of causative infectious agents in ulceration of the non-glandular part of the porcine stomach (pars oesophagea). In total, 150 stomachs from slaughter pigs were included, 75 from pigs that received a meal feed, 75 from pigs that received an equivalent pelleted feed with a smaller particle size. The pars oesophagea was macroscopically examined after slaughter. (q)PCR assays for H. suis, F. gastrosuis and H. pylori-like organisms were performed, as well as 16S rRNA sequencing for pars oesophagea microbiome analyses. All 150 pig stomachs showed lesions. F. gastrosuis was detected in 115 cases (77%) and H. suis in 117 cases (78%), with 92 cases (61%) of co-infection; H. pylori-like organisms were detected in one case. Higher infectious loads of H. suis increased the odds of severe gastric lesions (OR = 1.14, p = 0.038), while the presence of H. suis infection in the pyloric gland zone increased the probability of pars oesophageal erosions [16.4% (95% CI 0.6-32.2%)]. The causal effect of H. suis was mediated by decreased pars oesophageal microbiome diversity [-1.9% (95% CI - 5.0-1.2%)], increased abundances of Veillonella and Campylobacter spp., and decreased abundances of Lactobacillus, Escherichia-Shigella, and Enterobacteriaceae spp. Higher infectious loads of F. gastrosuis in the pars oesophagea decreased the odds of severe gastric lesions (OR = 0.8, p = 0.0014). Feed pelleting had no significant impact on the prevalence of severe gastric lesions (OR = 1.72, p = 0.28). H. suis infections are a risk factor for ulceration of the porcine pars oesophagea, probably mediated through alterations in pars oesophageal microbiome diversity and composition., (© 2024. The Author(s).)

Helicobacter pylori is a well-known pathogen that causes chronic gastritis, leading to the development of gastric cancer. This bacterium has also been detected in dogs, and symptoms similar to those in humans have been reported. The cytotoxin-associated gene A (CagA) is involved in pathogenesis through aberrant activation of host signal transduction, including the nuclear factor-kappa B (NF-κB) pathway. We have previously shown the anti-inflammatory effect of the G-protein-coupled estrogen receptor (GPER) via inhibiting of NF-κB activation in several cells. Therefore, here, we investigated the effect of GPER on CagA-mediated NF-κB promoter activity and showed that CagA overexpression in gastric cancer cells activated the NF-κB reporter and induced interleukin 8 (il-8) expression, both of which were inhibited by the GPER agonist.

Background: Helicobacter pylori is a worldwide pathogen that affects both animals and humans with a wide environmental distribution, causing serious health problems in humans. This research has timely addressed the topic of new sources of H. pylori infection, which is currently a global issue, especially in developing countries. For this purpose, 115 Tilapia fish, 50 freshwater samples, and 88 fish-handlers' stool samples were investigated for the presence of H. pylori in Qena Governorate, Egypt. The applied techniques were antigen screening tests, culturing, and molecular methods through ureC gene amplification, and 16 S rRNA characterization., Results: Helicobacter pylori was detected in 7.83%, 14%, 4.35%, and 12% of the investigated fish and water samples by culture and PCR methods, respectively. Out of the total studied participants, 40 tested positive for H. pylori when screened by stool antigen test, of which 35 (39.77%), and 31 (35.23%) were confirmed by conventional and molecular techniques, respectively. The Fisher's exact test has shown a statistically significant correlation between H. pylori infection, sex, and age as risk factors, while the association was insignificant concerning the residence. Males contracted the infection at a higher rate than females (48.08% and 16.67%, respectively). Also, H. pylori infection rate was the highest among fish-handlers aged 36-45 years old (46.67%), followed by the 26-35 years old age group (39.53%). With regard to the residence, a higher occurrence rate was recorded in the rural (36.07%) than the urban population (33.33%). Helicobacter pylori isolates harbored the highest antimicrobial resistance against ampicillin (100%), metronidazole (95.24%), while the least antimicrobial resistance was recorded against levofloxacin (21.43%), and clarithromycin (26.20%). The phylogenetic analysis revealed a high degree of homology between the isolates selected from Tilapia fish, freshwater, and fish-handlers., Conclusions: Our data emphasized the role that fish and freshwater play in disseminating H. pylori infection as one of the diseases that has a significant public health issue., (© 2023. The Author(s).)

Helicobacter pylori (H. pylori) is a zoonotic gastric microorganism capable of efficient interspecies transmission. Domesticated companion animals, particularly dogs and cats, serve as natural reservoirs for H. pylori. This phenomenon facilitates the extensive dissemination of H. pylori among households with pets. Hence, the prompt and precise identification of H. pylori in companion animals holds paramount importance for the well-being of both animals and their owners. With the assistance of Multienzyme Isothermal Rapid Amplification (MIRA) and CRISPR-Cas12a system, we successfully crafted a highly adaptable optical detection platform for H. pylori. Three sensor systems with corresponding visual interpretations were proposed. This study demonstrated a rapid turnaround time of approximately 45 min from DNA extraction to the result display. Moreover, this platform topped germiculture and real-time PCR in terms of sensitivity or efficiency in clinical diagnoses of 66 samples. This platform possesses significant potential as a versatile approach and represents the premiere application of CRISPR for the non-invasive detection of H. pylori in companion animals, thereby mitigating the dissemination of H. pylori among household members., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

While seven gastric non-Helicobacter pylori Helicobacter (NHPH) species are known to commonly colonize the stomach of cats and dogs, the potential of H. pylori and H. pylori-like organisms to infect animals remains controversial and was investigated in this study using gastric samples of 20 cats and 27 dogs. A Helicobacter genus-specific 16 S rRNA PCR assay, H. pylori-specific ureAB and glmM PCR assays and a nested PCR detecting 23 S rRNA in a Helicobacter genus-specific manner in a first round of PCR and a H. pylori-specific manner in a second round, were performed in combination with sequencing. Histopathological and anti-Helicobacter immunohistochemical evaluations were also performed. Based on 16 S rRNA sequence analysis, 39/47 animals (83%) appeared infected with canine/feline gastric NHPHs in the corpus and/or antrum. H. pylori-specific ureAB amplicons were obtained in samples of 22 stomachs (47%). One canine antrum sample positive in the ureAB assay was also positive in the H. pylori-specific glmM assay. While 36/47 (77%) animals had a positive sample in the first round of the nested 23 S rRNA PCR assay, all samples were negative in the second round. Sequence analysis of obtained amplicons and immunohistochemistry point towards the presence of unidentified H. pylori-like organisms in cats and dogs. Histopathological examination suggests a low pathogenic significance of the gastric Helicobacter spp. present in these animals. In conclusion, cats and dogs may be (co-)infected with gastric Helicobacter organisms other than the known gastric NHPHs. Culture and isolation should be performed to confirm this hypothesis., (© 2023. L’Institut National de Recherche en Agriculture, Alimentation et Environnement (INRAE).)

Gastritis and gastric ulcers are well-recognized conditions in cetaceans; bacteria of the genus Helicobacter are considered the primary cause of these diseases. Dolphins have been shown to be susceptible to infection by at least 2 gastric species of Helicobacter, H. cetorum and H. delphinicola, both of which are closely related to the human pathogen H. pylori. In the present study, we evaluated the carriage rate and relationship to gastric disease of H. cetorum and H. delphinicola, based on a study population of 82 dolphins maintained at 21 facilities in Japan. Of these 82 dolphins, 79 (96.3%) and 45 (54.9%) were positive for H. cetorum and H. delphinicola, respectively; H. delphinicola infection was significantly associated with chronic gastric diseases (odds rate: 5.9; 95% CI: 2.1-16.9), but no such association was detected for H. cetorum. Of the 21 facilities, 20 (95%) and 11 (55%) housed H. cetorum- and H. delphinicola-positive dolphins, respectively, and our study suggested that the transmission between dolphins occurs quickly within pools. These findings indicate that methods will need to be established to prevent the transmission of Helicobacter infections within facilities housing dolphins.

We report the isolation of Helicobacter ailurogastricus, a Helicobacter species that infects cats and dogs, from a person with multiple refractory gastric ulcers. In addition to H. suis, which infects pigs, Helicobacter species that infect cats and dogs should be considered as potential gastric pathogens in humans.

A 3.5-year-old female cheetah (Acinonyx jubatus) died after a 10-day history of anorexia, regurgitation and diarrhoea despite symptomatic therapy. At gross post-mortem examination, the stomach was blood-filled with mucosal thickening and multifocal ulcerations. The intestinal mucosa was thickened and reddened, and the intestinal lumen was filled with dark red to black pasty content. Gastric histological lesions were compatible with gastritis due to Helicobacter infection, which was confirmed by polymerase chain reaction. Histology of the intestines revealed a severe necrotizing neutrophilic enterocolitis with abundant intralesional curved to spiral bacteria, corresponding to Campylobacter jejuni, which were subsequently isolated from both small and large intestinal contents. No other intestinal pathogens were detected despite thorough investigations. These findings suggest that C. jejuni may have played an aetiological role in the enterocolitis. Such an association has not been previously reported in non-domestic felids., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: High frequency of Helicobacter pylori infection and the unknown mode of transmission prompted us to investigate H. pylori -wild housefly relationship. H. pylori causes chronic gastritis, peptic ulcers, and stomach cancer. H. pylori persists in the gut of the experimentally infected houseflies. The existence of H. pylori strains isolated from wild houseflies, on the other hand, has never been documented. Materials and Methods: In this study, 902 wild houseflies from different sites were identified as Musca domestica , then 60 flies were screened by traditional microbiological techniques and H. pylori -specific 16S rRNA gene. The antibiotic resistance (ART) was investigated phenotypically. Wild housefly gut bacterial isolates were further evaluated genotypically to have 23S rRNA gene mutation related to clarithromycin resistance. To find efficient therapeutic alternatives, the potency of three plant extracts (garlic, ginger, and lemon) and the wasp, Vespa orientalis venom was evaluated against H. pylori . The cytotoxic effect of the crude wasp venom, the most potent extract, against Vero and Colon cancer (Caco2) cell lines was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: All isolates from houseflies were positive. The isolated bacteria have variable resistance to frequently used antibiotics in all isolates. Minimum inhibitory concentration values of 15.625 mg/mL for both ginger and lemon extracts, 7.8125 mg/mL for garlic extract, and 0.0313 mg/mL for wasp venom were recorded. Wasp venom has the most potent antibacterial activity compared with the four antibiotics that are currently used in therapies against H. pylori . Conclusion: We conclude that wild houseflies can play a role in disseminating H. pylori. The housefly gut may be a suitable environment for the horizontal transfer of ART genes among its associated microbiome and H. pylori. Wasp venom proved its potential activity as a new and effective anti- H. pylori drug for both therapeutic and preventative usage.

Background: Helicobacter pylori is one of the most common bacterial infections and is widespread globally. It causes a variety of gastrointestinal disorders, though a great proportion of infections are asymptomatic. A total of 143 fresh stool samples were collected from apparently healthy farm and pet animals (43 cattle, 50 buffaloes, 50 sheep, 50 dogs, and 50 cats), in addition to 768 human stool samples. The samples were examined using stool antigen and rapid antibody tests, and further confirmation of glmM "human antigen-positive samples and animal milk samples" was conducted by polymerase chain reaction (PCR)., Results: The prevalence rates of H. pylori infection in animals were 22.2% and 16% in antibody and stool antigen tests, respectively. The detection rates were 28%, 24%, 12%, 10%, and 4.7% in cats, dogs, buffaloes, sheep, and cattle, respectively. On the other hand, the prevalence rate of H. pylori infection in human stool samples was 74.8%, and a statistically significant association was observed between prevalence and several factors, such as sex, age, and locality. PCR was performed to detect the glmM gene of H. pylori, and this gene was found in 21 of 27 human antigen-positive samples and 5 of 13 animal milk samples., Conclusions: H. pylori was detected in both human and animal samples. Furthermore, glmM was found in milk and human samples. Our findings suggest that pet and farm animals could transmit H. pylori infection to humans., (© 2023. The Author(s).)

Besides Helicobacter pylori, a Gram-negative bacterium that may cause gastric disorders in humans, non-Helicobacter pylori helicobacters (NHPH) may also colonize the stomach of humans and animals. In pigs, H. suis can induce gastritis and may play a role in gastric ulcer disease, possibly in association with Fusobacterium gastrosuis. In the present study, gastric samples from 71 slaughtered pigs and 14 hunted free range wild boars were tested for the presence of DNA of F. gastrosuis and gastric Helicobacter species associated with pigs, dogs cats and humans, using species-specific PCR assays, followed by sequencing of the amplicon. These gastric samples were also histopathologically evaluated. Almost all the pigs presented gastritis (95.8%). Helicobacter spp. were detected in 78.9% and F. gastrosuis in 35.2% of the animals. H. suis was the most frequently identified Helicobacter species (57.7% of the animals), followed by a H. pylori-like species (50.7%) and less often H. salomonis and H. felis (each in 2.8% of the animals). H. suis was most often detected in the glandular (distal) part of the stomach (pars oesophagea 9.9%, oxyntic mucosa 35.2%, antral mucosa 40.8%), while the H. pylori-like species was mainly found in the non-glandular (proximal) part of the stomach (pars oesophagea 39.4%, oxyntic mucosa 14.1%, antral mucosa 4.2%). The great majority of wild boars were also affected with gastritis (71.4%) and Helicobacter spp. and F. gastrosuis were detected in 64.3% and 42.9% of the animals, respectively. H. bizzozeronii and H. salomonis were the most frequently detected Helicobacter species, while a H. pylori-like species and H. suis were only occasionally identified. These findings suggest that these microorganisms can colonize the stomach of both porcine species and may be associated with gastric pathology. This should, however, be confirmed through bacterial isolation. This is the first description of the presence of F. gastrosuis DNA in the stomach of wild boars and a H. pylori-like species in the pars oesophagea of the porcine stomach., (© 2022. The Author(s).)

Helicobacter pullorum (H. pullorum) is a bacterium that colonizes the intestines of poultry and causes gastroenteritis. Because these species are known as human and/or animal pathogens, identification of H. pullorum is becoming increasingly necessary. The bacterium has been linked to colitis and hepatitis in humans after being transmitted by infected meat consumption. Misdiagnosis of other enteric zoonotic pathogens such as Campylobacter and other Helicobacter species makes the diagnosis of H. pullorum extremely difficult. This study focused on the molecular detection of H. pullorum from the stomach (proventriculus and gizzard) of different avian species as new target organs for detection and transmission between avian species. Proventriculus and gizzards were obtained from 40 freshly dead chickens and resident wild birds (n=40). Diarrhea was found in the farms that were surveyed. DNA was extracted from all collected samples to conduct PCR amplification. The samples were screened for Helicobacter genus-specific 16s using C97 and C05 primers. To confirm the existence of H. pullorum, the positive samples were sequenced. H. pullorum was recorded in two out of 40 chicken samples. In addition, H. pullorum was recorded in one out of 40 resident wild birds. The 16S rRNA gene sequence for Helicobacter genus-specific in poultry and wild birds showed a 100% homology. In conclusion, broiler chickens and resident wild birds are possible reservoirs for H. pullorum, according to this report, and possibly act as a source of infection for humans via the food supply., (Copyright© by the Polish Academy of Sciences.)

This article focuses on the pathogenic significance of Helicobacter species naturally colonizing the stomach of dogs, cats and pigs. These gastric "non-Helicobacter (H.) pylori Helicobacter species" (NHPH) are less well-known than the human adapted H. pylori. Helicobacter suis has been associated with gastritis and decreased daily weight gain in pigs. Several studies also attribute a role to this pathogen in the development of hyperkeratosis and ulceration of the non-glandular stratified squamous epithelium of the pars oesophagea of the porcine stomach. The stomach of dogs and cats can be colonized by several Helicobacter species but their pathogenic significance for these animals is probably low. Helicobacter suis as well as several canine and feline gastric Helicobacter species may also infect humans, resulting in gastritis, peptic and duodenal ulcers, and low-grade mucosa-associated lymphoid tissue lymphoma. These agents may be transmitted to humans most likely through direct or indirect contact with dogs, cats and pigs. Additional possible transmission routes include consumption of water and, for H. suis, also consumption of contaminated pork. It has been described that standard H. pylori eradication therapy is usually also effective to eradicate the NHPH in human patients, although acquired antimicrobial resistance may occasionally occur and porcine H. suis strains are intrinsically less susceptible to aminopenicillins than non-human primate H. suis strains and other gastric Helicobacter species. Virulence factors of H. suis and the canine and feline gastric Helicobacter species include urease activity, motility, chemotaxis, adhesins and gamma-glutamyl transpeptidase. These NHPH, however, lack orthologs of cytotoxin-associated gene pathogenicity island and vacuolating cytotoxin A, which are major virulence factors in H. pylori. It can be concluded that besides H. pylori, gastric Helicobacter species associated with dogs, cats and pigs are also clinically relevant in humans. Although recent research has provided better insights regarding pathogenic mechanisms and treatment strategies, a lot remains to be investigated, including true prevalence rates, exact modes of transmission and molecular pathways underlying disease development and progression., (© 2022. The Author(s).)

Background: Murine Helicobacter species have gained increasing awareness in mouse facilities over the last years. Infections with Helicobacter species may have an impact effect on the health of mice and might pose a zoonotic risk to researchers. To minimize the interference with experiments and hence contribute to the 3Rs, a reliable method of monitoring Helicobacter infections in animal facilities needs to be available. The aim of this study was to improve and validate the detection of the most common murine Helicobacter species., Material and Methods: A multiplex PCR assay was developed for identification of Helicobacter hepaticus, H. bilis, H. muridarum, H. rodentium, and H. typhlonius that could simultaneously detect these five strains in fecal samples. To ensure the quality of the results, the method was validated based on recommendations for in-house developed tests., Results: The method established was highly sensitive and specific. All five strains were detectable with a detection limit of 10 2 bacteria. Eight different mouse facilities were tested with the validated assay, and the following prevalence were found: H. rodentium 57%, H. hepaticus 46%, H. typhlonius 17%, H. bilis 12%, and H. muridarum 0%., Conclusion: The multiplex PCR is a reliable, economic, and time-saving diagnostic tool for routine health monitoring. Further prevalence studies are needed to confirm the high prevalence and hence importance of H. rodentium, as until now this agent is not yet listed in FELASA recommendations., (© 2022 John Wiley & Sons Ltd.)

Background: Companion animals like dogs play an important role in the lives of many people and are often considered to be members of families, but definitely, any contact with them poses an inherent risk of transmitting zoonotic pathogens. One of these pathogens is the genus Helicobacter which is linked to many disorders in human and animal., Objectives: The aim of this study was to investigate the presence of some zoonotic species of genus Helicobacter in companion dogs., Results: Through culturing in a special medium, nine samples (9%) were detected as infected (two pure and seven mixed culture). Based on multiplex-PCR, 13 samples (13%) were infected by Helicobacter spp. although none of them were infected by H. pylori. Species-specific PCR indicated that 38.5% or 5/13 of the samples were infected with H. heilmannii, while 15.45% or 2/13 of the samples were infected by H. billis. Multivariate logistic regression analysis showed that the age factor had a significant effect on Helicobacter spp. infection (odds ratio [OR] = 2.42, p = 0.01)., Conclusion: This study revealed the negligible faecal transmission of H. pylori. Moreover, due to the detection of H. Heilmannii and H. billis in feces and their association with human gastric diseases, dog owners should be educated about the risks and transmission modes of zoonotic bacterial infections of dogs., (© 2022 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Background: The relationships among gastric lymphoid follicular hyperplasia (GLFH), Helicobacter-like organisms (HLOs), and clinical signs have not been established in dogs., Objectives: To evaluate the epidemiologic, clinical, endoscopic, and histopathologic findings associated with GLFH in dogs, and determine the association of GLFH with HLOs and the French Bulldog (FB) breed., Animals: Two hundred eighty-eight dogs that underwent gastroscopy between 2013 and 2016., Methods: Retrospective, cross-sectional study. Gastric biopsy samples were reviewed and scored for inflammation and HLOs. Dogs were divided into 3 groups: group 1 (63 FBs), group 2 (45 non-FB brachycephalic dogs), and group 3 (180 nonbrachycephalic dogs). Variables were evaluated for their association with GLFH., Results: Univariate analysis determined that intact males, young age, vomiting, gastroscopic findings (discoloration, hemorrhage, and ulcers), and histopathologic findings (gastric lamina propria lymphocytic infiltration and HLO score) were associated with GLFH (P ≤ .03). In the multivariate analysis, GLFH was associated with the HLO score (odds ratio [OR] > 5 for HLO scores 1-2 and >15 for HLO score of 3; P < .001), with vomiting (OR > 4; P = .01) but not with FB breed (P = .76) and age (P = .1). The HLO score was associated with younger age (P < .001)., Conclusion and Clinical Importance: The HLO score was associated with a high GLFH score. Vomiting was associated with GLFH. Helicobacter-like organisms are highly prevalent in young dogs and GLFH is indirectly associated with this factor. Clinical relevance of the identification of GLFH and HLO remains to be determined., (© 2022 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Pathogens can affect physiological and immunological reactions in immunocompromised animals and genetically engineered mice. Specifically, murine norovirus (MNV), Helicobacter, and intestinal protozoa are prevalent in rodent laboratory facilities worldwide. In this study, microbiological test results of the soiled bedding of sentinel mice showed the prevalence of MNV (50.9%, 28/55), Helicobacter hepaticus (29.1%, 16/55), Trichomonas spp. (14.5%, 8/55), and Entamoeba spp. (32.7%, 18/55). No single infections were detected as all cases were confirmed to have complex infections with two or four pathogens. In previous studies, the success rate of the cross-fostering method was not perfect; therefore, in this study, the entire mouse strain of the SPF rodent facility was rederived using embryo transfer. For up to three years, we confirmed that the results were negative with regular health surveillance tests. Embryo transfer was, thus, determined to be an effective method for the rederivation of specific pathogen free (SPF) barrier mouse facilities. This is the report for the effectiveness of embryo transfer as an example of successful microbiological clean-up of a mouse colony with multiple infections in an entire SPF mouse facility and embryo transfer may be useful for rederiving.

Background: Neutrophil extracellular traps (NETs) represent a novel cellular mechanism of antimicrobial defense activity. Intravascular neutrophils produce extracellular web-like structures composed of chromatin, histones, and cytoplasmic granule proteins to attack and kill microbes. They may impact both pathogen and host; NETs correlate strongly with disseminated intravascular coagulation and mortality in critically ill humans. The mechanism was first discovered in human neutrophils in 2004. Presumptive heterophil extracellular traps (HETs) in a non-avian reptile species were first described in blood films of a gopher tortoise with systemic inflammation., Objective: While prior reports are limited to blood film review and in vitro studies, this descriptive case series highlights the cytological identification of presumptive HETs in nine reptile patients., Methods: Subjects included six gopher tortoises, one blood python (Python curtus ), one Burmese python ( P. bivittatus), and one desert king snake ( Lampropeltis getula splendida ). All six gopher tortoises ( Gopherus polyphemus ) had upper respiratory disease with bacterial etiology (including Helicobacter sp. and/or Mycoplasma sp.), and snakes had upper respiratory tract infection confirmed with serpentovirus (n = 2) or bacterial dermatitis (n = 1)., Results: Cytology samples with identified HETs included tissue imprints (n = 4), nasal discharge (n = 3), an oral swab (n = 1), and a fine needle aspirate of a skin lesion (n = 1). The identification of specific bacterial (n = 6) and/or viral pathogens (n = 2) was notable., Clinical Relevance: To the authors' knowledge, this is the first report of presumptive HETs recognized in reptile cytology specimens, suggesting an active cellular process in vivo in response to systemic inflammation in non-avian reptiles, and contributing to further understanding of extracellular traps in these species.

Helicobacter species have been reported in animals, some of which are of zoonotic importance. This study aimed to detect Helicobacter species among human and animal samples using conventional PCR assays and to identify their zoonotic potentials. Helicobacter species was identified in human and animal samples by genus-specific PCR assays and phylogenetic analysis of partial sequencing of the 16S ribosomal RNA gene. The results revealed that Helicobacter species DNA was detected in 13 of 29 (44.83%) of the human samples. H. pylori was identified in 2 (15.38%), and H. bovis was detected in 4 (30.77%), whereas 7 (53.85%) were unidentified. H. bovis and H. heilmannii were prevalent among the animal samples. Phylogenetic analysis revealed bootstrapping of sequences with H. cinaedi in camel, H. rappini in sheep and humans, and Wollinella succinogenes in humans. In conclusion, the occurrence of non-H. pylori infections among human and animal samples suggested zoonotic potentials., (Copyright© by the Polish Academy of Sciences.)

Background: Whereas non-Helicobacter pylori helicobacters, which are frequently detected in the stomachs of dogs and cats as a source of zoonoses, have attracted considerable attention, the role of pets in H. pylori epidemiology is unclear. In our previous study, an H. pylori infection was detected in the stomach of a dog (Dog 1). Here, we investigated the H. pylori infection status in the female offspring of Dog 1 (Dog 2) and its owner within the same household., Materials and Methods: Biopsy specimens were obtained from the dog's owner and tested for H. pylori. DNA from gastric biopsy samples of Dog 1, gastric fluid sediment of Dog 2, and bacteria from the stomach of the owner was obtained, and Helicobacter genus- and species-specific PCRs were performed. Then, sequence analyses of the partial region of the ureAB gene were conducted., Results: Samples from both dogs and the owner reacted positively in the genus-specific PCR and negative in the Helicobacter felis-, Helicobacter bizzozeronii-, and Helicobacter heilmannii sensu stricto-specific PCRs. All three samples also reacted positively in the H. pylori-specific PCR. Sequences of the partial ureAB gene from all subjects were identical., Conclusions: The results suggested that the two dogs and their owner were infected with an identical H. pylori strain. This report is the first to demonstrate that H. pylori can be transmitted between humans and dogs. Further studies are required to investigate the risk factors for the transmission of H. pylori between humans and dogs from the perspective of preventive epidemiology., (© 2021 John Wiley & Sons Ltd.)

Enterohepatic Helicobacter (EHH) species have been increasingly associated with acute gastroenteritis, inflammatory bowel disease and hepatobiliary diseases in humans. However, their host range and transmission routes are poorly understood. Therefore, the aim of this study was to determine the presence of EHH in healthy dogs using both cultivation-dependent and -independent methods. Three hundred and ninety faecal samples from domestic dogs without gastrointestinal symptoms were analysed between June 2018 and July 2019 in Valdivia (South of Chile). Samples were inoculated on selective medium and in parallel were filtrated over an antibiotic-free blood agar. Both media were incubated in a microaerobic atmosphere at 37°C for 7 days. Colonies were identified by PCR and phylogenetic analysis. A subset of 50 samples (half of them positive for EHH by cultivation and the remaining half negative) was analysed by PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) for direct detection. Cultivation method detected EHH in 15.4% (60/390) of the samples, being the most prevalent species H. canis (5.8%, 23/390) and H. canicola (5.1%, 20/390), followed by H. bilis (3.6%, 14/390) and 'H. winghamensis' (1.3%, 5/390). In contrast, PCR-DGGE method detected Helicobacter DNA in almost all (96%, 48/50) tested samples. On the other hand, the method used also allowed to isolate other Campylobacterales, in fact 44.3% (173/390) of the samples were positive for Campylobacter upsaliensis (43.3%, 169/390) followed by C. jejuni (2.0%, 8/390). Moreover, two strains that presented Campylobacter-like morphology were finally identified as Anaerobiospirillum succiniciproducens. Our results indicate that healthy domestic dogs commonly carry EHH and other Campylobacter species. However, further studies are needed to determine whether and how these Helicobacter and Campylobacter species can be transmitted to humans., (© 2021 Wiley-VCH GmbH.)

Background: Ménétrier-like disease is a rare hypertrophic canine gastropathy, reported in only seven dogs. Clinical signs are vomiting, anorexia and weight loss. Macroscopically, giant cerebriform gastric mucosal folds are typically seen in the corpus and fundus of the stomach. Histopathologically, fundic mucous cell hyperplasia and loss of parietal and chief cells are typical., Case Presentation: A nine-year-old spayed female Pointer had a history of intermittent vomiting, marked weight loss and hypoalbuminaemia. A gastroduodenoscopy was performed three times within three months with macroscopic changes remaining the same. The gastric mucosa of the corpus, fundus and proximal antrum was markedly irregular, with cerebriform mucosal folds. In the first gastric biopsies, histopathology revealed a moderate granulomatous gastritis, with a severe manifestation of Helicobacter-like organisms. Treatment for Helicobacter spp. decreased the vomiting slightly. The dog was diagnosed with concurrent leishmaniosis; the conventional anti-Leishmania treatment decreased the vomiting moderately, the hypoalbuminaemia resolved and the dog gained weight back to a normal body condition. Granulomatous gastritis was not present in the gastric biopsies after these treatments. The dog increased vomiting when palliative treatment (maropitant citrate, ondansetron and esomeprazole) was discontinued, and thus, full-thickness biopsies of the stomach were taken and Ménétrier-like disease was diagnosed. The affected area was too large to be surgically removed; thus, palliative treatment was reinstated. The dog remained clinically well 39 months after the first clinical presentation., Conclusions: This is the first report of Ménétrier-like disease in a dog with a simultaneous manifestation of granulomatous gastritis, helicobacteriosis and leishmaniosis. The clinical signs decreased after treatment of helicobacteriosis and leishmaniosis, but vomiting remained probably as a sign of Ménétrier-like disease. Treatment options for dogs are surgical removal of the abnormal area or palliative treatment. In humans, promising results for a cure have been shown with cetuximab treatment, a human monoclonal antibody, but no canine antibody is commercially available yet. The dog here was doing well 39 months after first presentation, which is the longest reported survival time for Ménétrier-like disease with only palliative treatment in dogs. Full-thickness biopsies are preferred in macroscopic hypertrophic lesions of the stomach for better assessment of Ménétrier-like disease.

Introduction. Helicobacter suis ( Helicobacter heilmannii type 1) commonly infects nonhuman primates but its clinical importance is in question. Aim. To characterize H. suis infection in a colony of rhesus macaques ( Macaca mulatta ) used in cognitive neuroscience research. Hypothesis/Gap Statement. Inquiries into the nature of Helicobacter suis in nonhuman primates are required to further define the organism's virulence and the experimental animal's gastric microbiome. Methodology. Animals with and without clinical signs of vomiting and abdominal pain ( n =5 and n =16, respectively) were evaluated by histology, culture, PCR amplification and sequencing, fluorescent in situ hybridization (FISH) and serology. Three of the five animals with clinical signs, an index case and two others, were evaluated before and after antimicrobial therapy. Results. The index animal had endoscopically visible ulcers and multifocal, moderate, chronic lymphoplasmacytic gastritis with intraglandular and luminal spiral bacteria. Antimicrobial therapy in the index animal achieved histologic improvement, elimination of endoscopically visible ulcers, and evident eradication but clinical signs persisted. In the other treated animals, gastritis scores were not consistently altered, gastric bacteria persisted, but vomiting and abdominal discomfort abated.Nineteen of 21 animals were PCR positive for H. suis and five animals were also PCR positive for H. pylori . Organisms were detected by FISH in 17 of 21 animals: 16S rRNA sequences of two of these were shown to be H. suis . Mild to moderate lymphoplasmacytic gastritis was seen in antrum, body and cardia, with antral gastritis more likely to be moderate than that of the body. Conclusion. No clear association between the bacterial numbers of Helicobacter spp. and the degree of inflammation was observed. H. suis is prevalent in this colony of Macaca mulatta but its clinical importance remains unclear. This study corroborates many of the findings in earlier studies of H. suis infection in macaques but also identifies at least one animal in which gastritis and endoscopically visible gastric ulcers were strongly associated with H. suis infection. In this study, serology was an inadequate biomarker for endoscopic evaluation in diagnosis of H. suis infection.

The binuclear metalloenzyme Helicobacter pylori arginase is important for pathogenesis of the bacterium in the human stomach. Despite conservation of the catalytic residues, this single Trp enzyme has an insertion sequence (-153ESEEKAWQKLCSL165-) that is extremely crucial to function. This sequence contains the critical residues, which are conserved in the homolog of other Helicobacter gastric pathogens. However, the underlying basis for the role of this motif in catalytic function is not completely understood. Here, we used biochemical, biophysical and molecular dynamics simulations studies to determine that Glu155 of this stretch interacts with both Lys57 and Ser152. These interactions are essential for positioning of the motif through Trp159, which is located near Glu155 (His122-Trp159-Tyr125 contact is essential to tertiary structural integrity). The individual or double mutation of Lys57 and Ser152 to Ala considerably reduces catalytic activity with Lys57 to Ala being more significant, indicating they are crucial to function. Our data suggest that the Lys57-Glu155-Ser152 interaction influences the positioning of the loop containing the catalytic His133 so that this His can participate in catalysis, thereby providing a mechanistic understanding into the role of this motif in catalytic function. Lys57 was also found only in the arginases of other Helicobacter gastric pathogens. Based on the non-conserved motif, we found a new molecule, which specifically inhibits this enzyme. Thus, the present study not only provides a molecular basis into the role of this motif in function, but also offers an opportunity for the design of inhibitors with greater efficacy., (© 2021 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

Helicobacter pylori infection is one of the leading causes of several gastroduodenal diseases, such as gastritis, peptic ulcer, and gastric cancer. In fact, H. pylori eradication provides a preventive effect against the incidence of gastric cancer. Amoxicillin is a commonly used antibiotic for H. pylori eradication. However, due to its easy degradation by gastric acid, it is necessary to administer it in a large dosage and to combine it with other antibiotics. This complexity and the strong side effects of H. pylori eradication therapy often lead to treatment failure. In this study, the chitosan/poly (acrylic acid) particles co-loaded with superparamagnetic iron oxide nanoparticles and amoxicillin (SPIO/AMO@PAA/CHI) are used as drug nano-carriers for H. pylori eradication therapy. In vitro and in vivo results show that the designed SPIO/AMO@PAA/CHI nanoparticles are biocompatible and could retain the biofilm inhibition and the bactericidal effect of amoxicillin against H. pylori . Moreover, the mucoadhesive property of chitosan allows SPIO/AMO@PAA/CHI nanoparticles to adhere to the gastric mucus layer and rapidly pass through the mucus layer after exposure to a magnetic field. When PAA is added, it competes with amoxicillin for chitosan, so that amoxicillin is quickly and continuously released between the mucus layer and the gastric epithelium and directly acts on H. pylori . Consequently, the use of this nano-carrier can extend the drug residence time in the stomach, reducing the drug dose and treatment period of H. pylori eradication therapy.

Objectives: The study was designed to determine the presence of Helicobacter genus and three species of H. pylori, H. bilis, and H. canis, in the duodenum, ileum, colon, and liver of stray cats. Moreover, the histopathological and immunohistochemical analyses have been performed., Methods: Samples were taken from the duodenum, ileum, colon, and liver of 30 cats for molecular and histopathological evaluations. Polymerase chain reaction was carried out for the detection of the Helicobacter genus in the mentioned samples. Then, species-specific primers were used in Helicobacter-positive samples., Results: Helicobacter genus prevalence rates in the duodenum, ileum, colon, and liver samples were 50%, 60%, 50%, and 43.3%, respectively. Helicobacter pylori, H. canis, and H. bilis were isolated from at least one tissue of 18 (60%), 13 (43.3%), and 8 (26.7%) of the cats, respectively. Immunohistochemical findings confirmed the presence of bacteria in the intestinal crypt or the mucosal layer of duodenum, ileum, colon, and hepatic sinusoids., Conclusion: In the present study, the concurrent infection of duodenum and liver was noticeable. Furthermore, the high prevalence of H. pylori in cats, as a well-known human pathogen, should be considered. High incidence of Helicobacter in gut and liver of Ahvaz stray cats is noticeable. According to the zoonotic importance of Helicobacter, more studies in the field of treatment and prevention are highly recommended.

Six Helicobacter -like isolates were recovered from 15 gastric mucosa samples of red foxes ( Vulpes vulpes ) shot by hunters in the surroundings of Ljubljana, Slovenia. Gram-negative, tightly coiled, intensely motile, 7-15 µm long and ≤1 µm wide bacteria grew on the biphasic blood agar plates. By using a genus-specific polymerase chain reaction (PCR), all isolates were confirmed as Helicobacter sp. and subsequently subjected to whole-genome sequencing (WGS). Five isolates showed a genome-wide average nucleotide identity (ANI) value of <95 % to the previously described Helicobacter species and one isolate was classified as Helicobacter felis . In the five unidentified isolates, the 16S rRNA gene sequence similarity to the type strains of all Helicobacter species ranged from 98.6 to 98.9 %. Their taxonomic status was established using a polyphasic taxonomic approach comprising the core genome-based phylogeny, morphological and phenotypic characteristics, including an analysis of matrix-assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectra. Phylogeny revealed the existence of three novel and well-supported clusters, with Helicobacter bizzozeronii and Helicobacter baculiformis being the most closely related species. The isolates also differed from the previously described species in their MALDI-TOF profiles and some biochemical characteristics. In conclusion, the data presented herein indicate that the obtained isolates, excluding H. felis isolate, represent three novel Helicobacter species, for which the names Helicobacter labacensis sp. nov., Helicobacter mehlei sp. nov., and Helicobacter vulpis sp. nov. are proposed, with isolates L9 T (=DSM 108823 T =CRBIP 111719 T ), L15 T (=DSM 108730 T =CCUG 72910 T ) and L2 T (=DSM 108727 T =CCUG 72909 T ) as type strains, respectively.

Pathogenic microbial detection and control in laboratory animal facilities is essential to guarantee animal welfare, data validity and reproducibility. Helicobacter spp. are known to affect mice health, what may interfere with experimental outcomes. This study aimed to screen for Helicobacter spp. in mice from animal facilities in Rio de Janeiro, Brazil using a PCR-based method. Primers designed to specifically identify Helicobacter spp. were used to amplify feces or intestine DNA extracted of mice from four different animal facilities. The expected 375 base pairs (bp) amplicon was purified, sequenced and a similarity of 95% was observed when compared to deposited sequences of H. hepaticus and H. bilis. In our screening, Helicobacter spp. was detected in ~59% of fecal and ~70% of intestine samples. Our study is the first to screen for Helicobacter spp. in mouse facilities of a Rio de Janeiro University using a low cost, rapid molecular diagnostic test. Although Helicobacter spp. screening is not mandatory according to Brazilian animal welfare regulation it is recommended by institutional animal health monitoring programs guidelines worldwide, including ARRIVE, AAALAC and FELASA.

Helicobacter pullorum is an emerging zoonotic pathogen that causes gastroenteritis in chickens and inflammatory bowel disease in humans ingesting contaminated meat. However, the mechanism by which the bacterium causes disease is unclear. Type six secretion system (T6SS) plays a major role in bacterial pathogenesis and adaptation. Haemolysin coregulated protein (Hcp) plays a central role in the structure of the T6SS pilus and acts as effector protein in certain bacteria. In this study, H. pullorum isolated from 156 caecal samples of broiler chickens was screened for the presence of T6SS Hcp gene via PCR amplification. 30.7% of caecal and 18.3% of liver samples tested positive for presence of H. pullorum . From these positive samples, 29.7% possessed the T6SS gene. In bacterial co-culture experiments, significant loss of viability (81.6-39.1%) was observed for H. pullorum -infected hepatocytes and presence of Hcp did not contribute to the loss of cell viability. Nevertheless, infection of erythrocytes with Hcp-positive isolates was associated with greater haemolytic activity compared to infection with Hcp-negative isolates. Therefore, presence of T6SS could be indicative of virulent strains meriting further studies to characterize this virulence factor in H. pullorum infection.

This article is a review of the most important, accessible, and relevant literature published between April 2018 and April 2019 in the field of Helicobacter species other than Helicobacter pylori. The initial part of the review covers new insights regarding the presence of gastric and enterohepatic non-H. pylori Helicobacter species (NHPH) in humans and animals, while the subsequent section focuses on the progress in our understanding of the pathogenicity and evolution of these species. Over the last year, relatively few cases of gastric NHPH infections in humans were published, with most NHPH infections being attributed to enterohepatic Helicobacters. A novel species, designated "Helicobacter caesarodunensis," was isolated from the blood of a febrile patient and numerous cases of human Helicobacter cinaedi infections underlined this species as a true emerging pathogen. With regard to NHPH in animals, canine/feline gastric NHPH cause little or no harm in their natural host; however they can become opportunistic when translocated to the hepatobiliary tract. The role of enterohepatic Helicobacter species in colorectal tumors in pets has also been highlighted. Several studies in rodent models have further elucidated the mechanisms underlying the development of NHPH-related disease, and the extra-gastric effects of a Helicobacter suis infection on brain homeostasis was also studied. Comparative genomics facilitated a breakthrough in the evolutionary history of Helicobacter in general and NHPH in particular. Investigation of the genome of Helicobacter apodemus revealed particular traits with regard to its virulence factors. A range of compounds including mulberries, dietary fiber, ginseng, and avian eggs which target the gut microbiota have also been shown to affect Helicobacter growth, with a potential therapeutic utilization and increase in survival., (© 2019 John Wiley & Sons Ltd.)

Helicobacter suis has been associated with development of gastric ulcers in the non-glandular part of the porcine stomach, possibly by affecting gastric acid secretion and altering the gastric microbiota. Fusobacterium gastrosuis is highly abundant in the gastric microbiota of H. suis-infected pigs and it was hypothesized that this micro-organism could play a role in the development of gastric ulceration. The aim of this study was to obtain further insights in the influence of a naturally acquired H. suis infection on the microbiota of the non-glandular part of the porcine stomach and in the pathogenic potential of F. gastrosuis. Infection with H. suis influenced the relative abundance of several taxa at phylum, family, genus and species level. H. suis-infected pigs showed a significantly higher colonization rate of F. gastrosuis in the non-glandular gastric region compared to non-infected pigs. In vitro, viable F. gastrosuis strains as well as their lysate induced death of both gastric and oesophageal epithelial cell lines. These gastric cell death inducing bacterial components were heat-labile. Genomic analysis revealed that genes are present in the F. gastrosuis genome with sequence similarity to genes described in other Fusobacterium spp. that encode factors involved in adhesion, invasion and induction of cell death as well as in immune evasion. We hypothesize that, in a gastric environment altered by H. suis, colonization and invasion of the non-glandular porcine stomach region and production of epithelial cell death inducing metabolites by F. gastrosuis, play a role in gastric ulceration.

Helicobacter infection has been associated with hepatobiliary diseases in humans and animals. The aims of this study were to identify Helicobacter species in the hepatobiliary tract of dogs and to elucidate the possible association of these bacteria in liver diseases. Twenty-seven gastric and hepatobiliary samples were collected from 33 dogs with hepatic lesions and 17 dogs with no liver histological changes. Warthin-Starry staining, immunohistochemical assay, and PCR were performed to detect the presence of Helicobacter. Helicobacter genus was detected in 21.2% of the samples with hepatic lesions. The main lesion was chronic hepatitis. Immunohistochemistry revealed infection in liver (1/5) and gallbladder (1/3) 32 samples. The sequence analysis of seven amplicons of the 16S rRNA gene of Helicobacter genus from hepatobiliary samples showed 97.8 to 100% of nucleotide identity with gastric helicobacter. One amplicon of the ureA and ureB gene of Helicobacter genus from the stomach showed 89.1 to 90.7% nucleotide identity with H. heilmannii. The presence of Helicobacter genus in liver samples showing hepatic lesions suggests the involvement of these bacteria in the etiology of hepatobiliary disease in dogs. DNA sequences were similar to gastric Helicobacter species, reinforcing the hypothesis of bacterial translocation from the stomach to liver by the biliary pathway.

Helicobacter suis colonizes the stomach of most pigs and is the most prevalent non-Helicobacter pylori Helicobacter species found in the human stomach. In the human host, H. suis contributes to the development of chronic gastritis, peptic ulcer disease and MALT lymphoma, whereas in pigs it is associated with gastritis, decreased growth and ulcers. Here, we demonstrate that the level of H. pylori and H. suis binding to human and pig gastric mucins varies between individuals with species dependent specificity. The binding optimum of H. pylori is at neutral pH whereas that of H. suis has an acidic pH optimum, and the mucins that H. pylori bind to are different than those that H. suis bind to. Mass spectrometric analysis of mucin O-glycans from the porcine mucin showed that individual variation in binding is reflected by a difference in glycosylation; of 109 oligosaccharide structures identified, only 14 were present in all examined samples. H. suis binding to mucins correlated with glycans containing sulfate, sialic acid and terminal galactose. Among the glycolipids present in pig stomach, binding to lactotetraosylceramide (Galβ3GlcNAcβ3Galβ4Glcβ1Cer) was identified, and adhesion to Galβ3GlcNAcβ3Galβ4Glc at both acidic and neutral pH was confirmed using other glycoconjugates. Together with that H. suis bound to DNA (used as a proxy for acidic charge), we conclude that H. suis has two binding modes: one to glycans terminating with Galβ3GlcNAc, and one to negatively charged structures. Identification of the glycan structures H. suis interacts with can contribute to development of therapeutic strategies alternative to antibiotics.

Helicobacter pylori cagA-positive strains are critically involved in the development of gastric cancer. Upon delivery into gastric epithelial cells via type IV secretion, the cagA-encoded CagA interacts with and thereby perturbs the pro-oncogenic phosphatase SHP2 and the polarity-regulating kinase PAR1b via the tyrosine-phosphorylated EPIYA-C/D segment and the CM sequence, respectively. Importantly, sequences spanning these binding regions exhibit variations among CagA proteins, which influence the pathobiological/oncogenic potential of individual CagA. Here we isolated an H. pylori strain (Hp&#95;TH2099) naturally infecting the stomach of a housed macaque, indicating a zoonotic feature of H. pylori infection. Whole genome sequence analysis revealed that Hp&#95;TH2099 belongs to the hpAsia2 cluster and possesses ABC-type Western CagA, which contains hitherto unreported variations in both EPIYA-C and CM sequences. The CM variations almost totally abolished PAR1b binding. Whereas pTyr + 5 variation in the EPIYA-C segment potentiated SHP2-binding affinity, pTyr-2 variation dampened CagA tyrosine phosphorylation and thus impeded CagA-SHP2 complex formation. As opposed to the H. pylori standard strain, infection of mouse ES cell-derived gastric organoids with Hp&#95;TH2099 failed to elicit CagA-dependent epithelial destruction. Thus, the macaque-isolated H. pylori showed low virulence due to attenuated CagA activity through multiple substitutions in the sequences involved in binding with SHP2 and PAR1b.

Since the discovery of the human pathogen Helicobacter pylori, various other Helicobacter species have been identified in the stomach of domesticated and wild mammals. To better understand the evolutionary history of these ecologically similar but genetically distinct species, we analyzed 108 gastric Helicobacter genomes and included 54 enterohepatic Helicobacter genomes for comparison purposes. An admixture analysis supported the presence of an ecological barrier, preventing the genetic exchange between the gastric and enterohepatic Helicobacter species, and unraveled many gene flow events within and across species residing in the stomach. As pets can be colonized by multiple gastric Helicobacter species, the genetic exchange between the canine and feline strains was evident, with H. heilmannii and H. bizzozeronii showing the highest interspecies recombination. An admixture between H. pylori (in particular, the ancestral African strains), H. acinonychis from wild felines and H. cetorum from marine mammals was also identified. Because these latter species do not share the same host, this phenomenon is most likely a remaining signal of shared ancestry. A reconstruction of the time of divergence of the gastric Helicobacter spp. revealed that the domestic animal-related Helicobacter species evolved in parallel with H. pylori and its two closest relatives (H. acinonychis and H. cetorum), rather than together.

Ulceration of the non-glandular part of the stomach is a common disease entity of pigs worldwide, with prevalences of up to 93%. It may result in decreased daily weight gain, decreased feed intake and sudden death, thus leading to significant economic losses, as well as animal welfare issues. The aetiology is multifactorial. Diet particle size, management and infectious agents have been hypothesised to be involved. The exact mechanism behind porcine gastric ulceration is, however, not completely clear. The aim of this article is to provide an overview of the role of infectious agents in the development of porcine gastric ulceration. Results of recent studies indicate that Helicobacter suis infection plays an important role in gastric ulceration, probably by affecting gastric acid secretion through alteration of the number and/or function of parietal, D and G cells. In a gastric environment altered by H. suis, higher numbers of Fusobacterium gastrosuis are present and this novel pathogen has a potential role in the development of porcine gastric ulceration. Inoculation of pigs with Lactobacillus sp., Bacillus sp. or Helicobacter pylori-like bacteria in combination with a carbohydrate-rich diet may induce gastric lesions. It has been hypothesised that production of short chain fatty acids by some of these bacteria might be involved in the pathogenesis of porcine gastric ulceration, but the lack of taxonomic characterisation hampers the interpretation of these studies. Severe infectious diseases have also been associated with gastric lesions, probably due to interruption in feed intake and/or histamine release. Other agents, including fungi and parasites such as Hyostrongylus rubidus and Ascaris suum, have occasionally been associated with gastric lesions in pigs., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Helicobacter and Campylobacter species (spp.) colonize the gastrointestinal tract of various domesticated animals. Apart from their pathogenic significance in animals, several species are of zoonotic importance as well. For most non-domesticated animal spp., however, little is known on the presence and importance of these agents. Therefore, we investigated the presence of Helicobacter and Campylobacter spp. in marine and terrestrial zoo mammals. Faecal samples of various marine and terrestrial zoo mammals were collected from 6 different zoos in Belgium. These samples were tested for the presence of Helicobacter and Campylobacter spp. by isolation and direct demonstration of DNA using genus-specific PCRs, followed by sequencing of the obtained amplicons. Helicobacter spp. were detected in 12 and Campylobacter spp. in 5 of the 22 faecal samples from marine mammals. In 4 of 5 dolphins, H. cetorum was demonstrated, a well-known pathogen associated with gastritis and gastric ulceration in marine mammals. C. insulaenigrae was isolated from 4 of 6 sea lions and from 1 of 11 seals. To our knowledge, this is the first description of the presence of C. insulaenigrae on the European mainland. Helicobacter spp. were detected in 5 and Campylobacter spp. (mainly C. jejuni subsp. jejuni and C. coli) in 9 of the 26 faecal samples from terrestrial mammals. Potential novel enterohepatic Helicobacter spp. were detected in both marine and terrestrial zoo mammals. For the first time, the presence of several known and unknown Campylobacter and Helicobacter spp. was demonstrated in the gastrointestinal tract of various marine and terrestrial zoo mammals. Further investigation is needed on the pathogenic and zoonotic importance of these species., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The minimum inhibitory concentration of bambermycin on three porcine Helicobacter suis strains was shown to be 8 μg/mL. The effect of in-feed medication with this antibiotic on the course of a gastric infection with one of these strains, the host response and the gastric microbiota was determined in mice, as all of these parameters may be involved in gastric pathology. In H. suis infected mice which were not treated with bambermycin, an increased number of infiltrating B-cells, T-cells and macrophages in combination with a Th2 response was demonstrated, as well as a decreased parietal cell mass. Compared to this non-treated, infected group, in H. suis infected mice medicated with bambermycin, gastric H. suis colonization was not altered, but a decreased number of infiltrating T-cells, B-cells and macrophages as well as downregulated expressions of IL-1β, IL-8M, IL-10 and IFN-γ were demonstrated and the parietal cell mass was not affected. In bambermycin treated mice that were not infected with H. suis, the number of infiltrating T-cells and expression of IL-1β were lower than in non-infected mice that did not receive bambermycin. Gastric microbiota analysis indicated that the relative abundance of bacteria that might exert unfavorable effects on the host was decreased during bambermycin supplementation. In conclusion, bambermycin did not affect H. suis colonization, but decreased gastric inflammation and inhibited the effects of a H. suis infection on parietal cell loss. Not only direct interaction of H. suis with parietal cells, but also inflammation may play a role in death of these gastric acid producing cells.

Helicobacter suis is the second most prevalent Helicobacter species in the stomach of humans suffering from gastric disease. This bacterium mainly inhabits the stomach of domesticated pigs, in which it causes gastric disease, but it appears to be absent in wild boars. Interestingly, it also colonizes the stomach of asymptomatic rhesus and cynomolgus monkeys. The origin of modern human-, pig- or non-human primate-associated H. suis strains in these respective host populations was hitherto unknown. Here we show that H. suis in pigs possibly originates from non-human primates. Our data suggest that a host jump from macaques to pigs happened between 100 000 and 15 000 years ago and that pig domestication has had a significant impact on the spread of H. suis in the pig population, from where this pathogen occasionally infects humans. Thus, in contrast to our expectations, H. suis appears to have evolved in its main host in a completely different way than its close relative Helicobacter pylori in humans.

OBJECTIVE To evaluate the correlation between the density of native gastric Helicobacter spp and the presence of gastric lesions in dogs. ANIMALS 80 dogs of various breeds, sexes, and ages. PROCEDURES Gastroscopic and histologic examinations were performed for all dogs. Helicobacter spp were detected by combining evaluation of urease activity and results of bacteriologic culture, microscopic observation, and a 16S rRNA PCR assay. The density of Helicobacter-like organisms was evaluated with light microscopy by use of Warthin-Starry modified stain. Correlations were evaluated by use of the Spearman correlation analysis. RESULTS Gastritis was found in 55 of 80 dogs and classified as mild (n = 31), moderate (16), or severe (8). Of these 55 dogs, only 8 had clinical signs. Histologic examination revealed some degree of lymphocytic-plasmacytic infiltrate, mild eosinophilia, and neutrophilic inflammation in the lamina propria. Seventy-six dogs had positive results for Helicobacter spp. Helicobacter pylori DNA was not detected. Low density and homogeneous distribution of Helicobacter spp were observed in all gastric zones. CONCLUSIONS AND CLINICAL RELEVANCE A significant correlation between density of Helicobacter spp and gastroscopic or histologic lesions was not detected. These findings supported the contention that there is no correlation between general Helicobacter spp density or numbers and gastritis in dogs.

Background: Serum gastrin concentration can help diagnose gastrinomas in dogs if >3-10× the upper reference limit (URL), but antisecretory therapy and other conditions can also cause hypergastrinemia. Effects of antisecretory therapy (famotidine or ranitidine, omeprazole) on serum gastrin concentration in dogs with chronic enteropathy (CE) and its biological variation (BV) are unknown. Aim of the study was to evaluate serum gastrin in acid-suppressant-treated or -naïve CE dogs; test the association between serum gastrin and histopathologic findings in acid-suppressant-naïve CE dogs; and evaluate the BV of serum gastrin in dogs not receiving any gastric acid suppressive therapy. Samples from 231 dogs were used and serum gastrin was measured by chemiluminescence assay. Gastric and duodenal histologic lesions were evaluated and graded. BV of serum gastrin was evaluated in serial samples., Results: Serum gastrin concentrations were significantly higher in acid-suppressant-treated than acid-suppressant-naïve dogs (P = 0.0245), with significantly higher concentrations in proton pump inhibitor (PPI)- than H 2 -antihistamine-treated patients (P = 0.0053). More PPI- than H 2 -antihistamine-treated dogs had gastrin concentrations above URL (P = 0.0205), but not >3× nor >10× the URL. Serum gastrin concentrations correlated with the severity of gastric antral epithelial injury (P = 0.0069) but not with any other lesions or the presence/numbers of spiral bacteria in gastric biopsies. Intra- and inter-individual BV were 43.4 and 21.6%, respectively, in acid-suppressant-naïve dogs, with a reciprocal individuality index of 0.49 and a critical difference of ≥29.5 ng/L., Conclusions: Antisecretory (particularly PPI) treatment leads to hypergastrinemia in CE dogs, but the concentrations seen in this study are unlikely to compromise a diagnosis of gastrinoma. Use of a population-based URL for canine serum gastrin and a URL of ≤27.8 ng/L are appropriate.

The aim of this study was to assess the suitability of invasive and non-invasive methods used to diagnose Helicobacter spp. in the stomachs of dogs. The study was carried out on 30 dogs of both sexes and different breeds, between one and 15 years old. A histopathologic examination, a microbiological culture, a rapid urease test, a direct bacteriological preparation and a nested PCR assay were carried out. Gastric Helicobacter spp. was identified in gastric biopsy specimens from 16 (53.3%) dogs using direct bacteriological preparation, in four (13.3%) dogs based on a culture, in 23 (76.6%) dogs using the rapid urease test and in 21 (70,0%) dogs based on a histopathological assessment of the biopsy specimens. The nested PCR of the gastric biopsy specimens revealed gastric Helicobacter spp. in all the dogs (100%). A saliva PCR assay revealed gastric Helicobacter spp. in 23 (76.6%) dogs, while stool PCR revealed the bacterium in seven (23.3%) dogs. We found that invasive methods were more accurate than non-invasive methods in detecting a Helicobacter spp. infection in dogs. In addition, the nested PCR method used to evaluate the gastric mucosal biopsy specimens was the most accurate test for detecting Helicobacter spp. It was further found that the PCR-based saliva assay was the best non-invasive method for detecting Helicobacter spp. However, taking into consideration that most of the diagnostic methods used to detect this bacterium have drawbacks, at least two diagnostic methods should be used to detect Helicobacter spp. as is done in human medicine.

The current article is a review of the most important and relevant literature published in 2016 and early 2017 on non-Helicobacter pylori Helicobacter infections in humans and animals, as well as interactions between H. pylori and the microbiota of the stomach and other organs. Some putative new Helicobacter species were identified in sea otters, wild boars, dogs, and mice. Many cases of Helicobacter fennelliae and Helicobacter cinaedi infection have been reported in humans, mostly in immunocompromised patients. Mouse models have been used frequently as a model to investigate human Helicobacter infection, although some studies have investigated the pathogenesis of Helicobacters in their natural host, as was the case for Helicobacter suis infection in pigs. Our understanding of both the gastric and gut microbiome has made progress and, in addition, interactions between H. pylori and the microbiome were demonstrated to go beyond the stomach. Some new approaches of preventing Helicobacter infection or its related pathologies were investigated and, in this respect, the probiotic properties of Saccharomyces, Lactobacillus and Bifidobacterium spp. were confirmed., (© 2017 John Wiley & Sons Ltd.)

Reptiles have been shown to host a significant Helicobacter diversity. In order to survive, reptile-associated Helicobacter lineages need to be adapted to the thermally dynamic environment encountered in a poikilothermic host. The whole genomes of reptile-associated Helicobacter lineages can provide insights in Helicobacter host adaptation and coevolution. These aspects were explored by comparing the genomes of reptile-, bird-, and mammal-associated Helicobacter lineages. Based on average nucleotide identity, all reptile-associated Helicobacter lineages in this study could be considered distinct species. A whole genome-based phylogeny showed two distinct clades, one associated with chelonians and one associated with lizards. The phylogeny indicates initial adaptation to an anatomical niche, which is followed by an ancient host jump and subsequent diversification. Furthermore, the ability to grow at low temperatures, which might reflect thermal adaptation to a reptilian host, originated at least twice in Helicobacter evolution. A putative tricarballylate catabolism locus was specifically present in Campylobacter and Helicobacter isolates from reptiles. The phylogeny of reptile-associated Helicobacter parallels host association, indicating a high level of host specificity. The high diversity and deep branching within these clades supports long-term coevolution with, and extensive radiation within the respective reptilian host type.

Epidemiological and pathological studies of Helicobacter spp. in canine stomachs in Japan were performed to investigate strain specific pathogenicity. Gastric biopsies from 144 dogs with gastrointestinal diseases were evaluated for the presence of Helicobacter spp. using genus and species specific PCRs for Helicobacter felis, Helicobacter bizzozeronii, Helicobacter heilmannii sensu stricto (s.s.) and Helicobacter pylori. PCR indicated that 50/144 (34.7%) dogs were infected with Helicobacter spp. Of the genus positive samples, 21/50 could not be amplified by any of the species specific PCRs. To investigate Helicobacter at the species level, partial ureAB gene sequences from 48/50 genus positive samples were determined; 47 strains were identified. Thirty-five strains from 45 cases were closely related to H. heilmannii s.s. (89-99% sequence similarity), seven strains from seven cases were closely related to H. bizzozeronii (95-99% sequence similarity), three strains from three cases were closely related to Helicobacter felis (86%, 98% and 99% sequence similarity), one strain from one case was closely related to Helicobacter salomonis (99% sequence similarity) and one strain from one case was closely related to H. pylori (99% sequence similarity). Dogs infected with Helicobacter spp. most similar to H. heilmannii s.s. had a higher frequency of moderate to severe gastritis than dogs negative for Helicobacter spp. (P=0.044). In conclusion, the predominant Helicobacter spp. detected in canine stomachs in our study were most closely related to H. heilmannii s.s. and displayed substantial genetic diversity. Infection with Helicobacter spp. may be associated with more severe gastritis in dogs., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Gastric mRNA expression of markers for acid secretion and inflammation and presence of gastric ulceration was studied in naturally Helicobacter suis-infected and non-infected 2-3 months old, 6-8 months old and adult pigs. In H. suis-infected 2-3 months old pigs, IL-8 and IL-1β transcript levels were upregulated in the pyloric gland zone, indicating an innate immune response. A similar response was demonstrated in the fundic gland zone of adult pigs, potentially due to a shift of H. suis colonization from the pyloric to the fundic gland zone. A Treg response in combination with decreased expressions of IL-8, IL-17A and IFN-γ was indicated to be present in the H. suis-infected 6-8 months old pigs, which may have contributed to persistence of H. suis. In H. suis-infected adult pigs, a Treg response accompanied by a Th17 response was indicated, which may have played a role in the decreased number of H. suis bacteria in the stomach of this age group. The decreased G-cell mass and upregulated expression of somatostatin indicated decreased acid secretion in H. suis-infected 6-8 months old pigs. In H. suis-infected adult pigs, upregulation of most markers for gastric acid secretion and increased G-cell mass was detected. Presence of severe hyperkeratosis and erosions in the non-glandular part of the stomach were mainly seen in the H. suis-positive groups. These results show that H. suis infection affects the expression of markers for acid secretion and inflammation and indicate that these effects differ depending on the infection phase.

Infections with Helicobacter spp. are known to have serious effects on rodent health, especially in immunocompromised animals. In this study three approaches were used to eradicate Helicobacter spp. infection in mice with a deficiency in intestinal proteoglycan (mucin2), namely triple oral antibiotic therapy (amoxicillin, clarithromycin and metronidazole), cross-fostering of neonatal pups by surrogate mothers negative for Helicobacter spp., and in vitro fertilization (IVF) with embryo transfer into Helicobacter-free mothers. However, triple antibiotic therapy in mice with mucin2 deficiency was not effective and had negative effects on reproductive performance, and high susceptibility of mucin2-deficient mice to Helicobacter spp. during the first 12 h after birth rendered cross-fostering impossible. Only IVF with embryo transfer was effective in eradicating Helicobacter infection from transgenic mice with mucin2 deficiency.