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.

Background Enterohepatic species of the genus Helicobacter (EHH) are emerging pathogens that have been associated with gastrointestinal and hepatobiliary diseases in humans. However, studies on their pathogenicity are scarce. Galleria mellonella is a recently proposed model for the study of virulence in different pathogens, such as Campylobacter spp. and Helicobacter pylori. Despite this, its usefulness in EHH has not yet been evaluated. Therefore, we determined the pathogenic potential of different EHH species isolated from dogs in this infection model. Materials and Methods Four species of EHH (H. bilis, H. canicola, H. canis, and 'H. winghamensis') isolated from fecal samples from domestic dogs were evaluated. Three strains of each species were inoculated in cohorts of G. mellonella at a concentration of 1 x 10(7) CFU/mL. Survival curves were determined by the Kaplan-Meier method. In addition, the quantification of melanin, bacterial load in hemolymph, and histopathology were evaluated daily post-infection (pi). Results G. mellonella larvae are susceptible to EHH infection, exhibiting intra- and inter-species variability. Melanin production became evident from 4 h pi and increased throughout the assay. All species were recovered from the hemolymph after 20 min pi; however, only H. canis could be recovered up to 48 h pi. Histopathology revealed cellular and humoral immune response, evidencing accumulation of hemocytes, nodulation, and melanin deposition in different tissues. Conclusion EHH species carried by dogs have considerable pathogenic potential, being H. canicola the species with the highest degree of virulence. Thus, G. mellonella is a useful model to assess virulence in these emerging pathogens.

The genus Helicobacter defined just over 30 years ago, is a highly diverse and fast-growing group of bacteria that are able to persistently colonize a wide range of animals. The members of this genus are subdivided into two groups with different ecological niches, associated pathologies, and phylogenetic relationships: the gastric Helicobacter (GH) and the enterohepatic Helicobacter (EHH) species. Although GH have been mostly studied, EHH species have become increasingly important as emerging human pathogens and potential zoonotic agents in the last years. This group of bacteria has been associated with the development of several diseases in humans from acute pathologies like gastroenteritis to chronic pathologies that include inflammatory bowel disease, and liver and gallbladder diseases. However, their reservoirs, as well as their routes of transmission, have not been well established yet. Therefore, this review summarizes the current knowledge of taxonomy, epidemiology, and clinical role of the EHH group.

Campylobacter jejuni is a major cause of acute gastroenteritis worldwide. However, it has also been associated with other diseases such as bacteremia and with several post-infection sequelae. Although campylobacteriosis is usually a self-limited infection, antibiotics are indicated for severe and chronic conditions. Unfortunately, several industrialised nations have reported a substantial increase in antibiotic resistance of C. jejuni. However, there is still a lack of knowledge about the epidemiology of resistance developed by this pathogen in the developing world. For this reason, our objective was to determine the resistance of clinical C. jejuni strains to ciprofloxacin and erythromycin in Chile and their associated genotypes.Fifty C. jejuni isolates recovered from fecal samples of people with acute gastroenteritis, in central and southern Chile between 2006 and 2015, were analysed. Resistance to erythromycin and ciprofloxacin was assessed by disk diffusion and agar dilution methods. Furthermore, these strains were genotyped by Multilocus Sequence Typing (MLST).Only one of the isolates was resistant to erythromycin. However, 48% of them were resistant to ciprofloxacin. The minimal inhibitory concentration of these ciprofloxacin-resistant isolates was in the range between 4 and 32 μg/ml. Moreover, MLST analyses showed that most ciprofloxacin-resistant strains were grouped into three dominant clonal complexes (ST-21, ST-48 and ST-353), while the unique strain resistant to both antibiotics belonged to the ST-45 complex. Our results evidence a high ciprofloxacin resistance and suggest that there is a dissemination of resistant clonal lineages responsible for cases of campylobacteriosis in Chile. Further studies should elucidate the origin of these resistant genotypes.

This work performed a phenotypic and genotypic characterization of 79 clinical isolates of Enterobacteriaceae and Pseudomonadaceae collected in hospitals of Southern Ecuadorin 2013. Our results showed a high incidence of β-lactamases and ESBLs with blaTEM and blaCTX-M as the prevalent genes, respectively. By direct sequencing of PCR amplicons, the different β-lactamases and variants of the genes were also distinguished. Our results revealed a predominance of TEM-1 β-lactamase and the presence of different CTX-M variants with a prevalence of CTX-M-15. Two infrequent CTX-M variants in South America were also identified. To the best of our knowledge, this is one of the first studies describing the genetic characteristics of β-lactamases in Ecuador.

The prevalence and antimicrobial behaviour of Campylobacter jejuni/coli isolated from backyard chickens from Southern Ecuador were determined. This study revealed that Campylobacter sp. frequency was 41.7% with high resistance to tetracycline (94.0%) and ciprofloxacin (88%). C. jejuni was the most frequently isolated species (32.5%), followed by C. coli (9.2%) being poultry colonization by Campylobacter not region dependent. This is the first study on Campylobacter antimicrobial resistance in backyard chickens in Southern Ecuador.

Background: Campylobacter jejuni and C. coli are important agents of diarrhea in human beings. Several animals are reservoirs for these bacteria and they may contribute to environmental contamination through fecal shedding. The presence of dog feces in urban settings could be a risk of Campylobacter environmental contamination and a problem of public health concern. Besides environmental contamination, the increasing antimicrobial resistance among Campylobacter is another issue of public health significance. The aims of this study were to determine the prevalence of C. jejuni and C. coli in dog feces collected from public parks of Loja city, Ecuador, and establishing their susceptibility to six antibiotics.Materials, Methods & Results: A total of 70 fresh-looking non-diarrheal dog fecal samples were collected from public parks of Loja city. An amount (c. a. 1 g) of each fecal specimen was collected with a swab, placed in a tube containing Bolton enrichment broth and processed within the next six hours. Campylobacter spp. were isolated by direct plating onto Butzler selective agar and into Bolton enrichment broth, which were incubated at 42 degrees C for 48 h in microaerophilic conditions. From Bolton enrichment broth subcultures to Butzler selective agar were made and also to blood agar, the latter using the passive filtration method in order to isolate C. upsaliensis. Identification was made firstly using the traditional phenotypic test (catalase, oxidase, oxygen tolerance, growth at 25 degrees C, hippurate and indoxyl acetate hydrolysis, and cephalothin and nalidixic acid susceptibility) and corroborated by the genus-specific polymerase chain reaction (PCR) and by the multiplex PCR (m-PCR) method proposed for the characterization of the six most common clinical Campylobacter species. Susceptibility to ampicillin, amoxicillin/clavulanic acid, erythromycin, tetracycline, ciprofloxacin and gentamycin was performed by the disk diffusion method following the 2014 recommendations of The European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Committee for the Antibiogram of the French Society of Microbiology. Campylobacter was isolated in 7/70 samples (10%) corresponding 4 (5.7%) to C. jejuni and 3 (4.3%) to C. coli. C. upsaliensis was not found. All the strains were susceptible to ampicillin, amoxicillin/clavulanic acid, erythromycin and gentamycin, six were resistant to tetracycline but all the seven strains were ciprofloxacin-resistant.Discussion: The isolation frequency of C. jejuni and C. coli in dog feces collected in the public parks of Loja city was higher than that reported by Wright (4.6%) for similar type of samples. However, it was lower than that shown in the other reports when dogs were directly sampled. Besides the aesthetically undesirable aspect of the presence of feces in public settings, they may represent a hygiene and public health problem because dog feces may contain potentially pathogenic bacteria for humans, like Salmonella spp., Escherichia coli, Yersinia spp. and Campylobacter spp. All seven isolated strains were susceptible to ampicillin, amoxicillin/clavulanic acid, erythromycin and gentamycin, six were resistant to tetracycline and surprisingly all of them were resistant to ciprofloxacin. The latter fact reflects the overuse of fluoroquinolones in animals, representing an emerging epidemiological problem with implications for treatment in humans. Dog feces deposited in public parks of Loja city yielded C. jejuni and C. coli suggesting that dogs could be a common source of environmental contamination with the additional risk of widespread ciprofloxacin resistant strains.