c Antimicrobial resistance in microbes poses a global and increasing threat to public health. The horizontal transfer of antimicrobial resistance genes was thought to be due largely to conjugative plasmids or transposons, with only a minor part being played by transduction through bacteriophages. However, whole-genome sequencing has recently shown that the latter mechanism could be highly important in the exchange of antimicrobial resistance genes between microorganisms and environments. The transfer of antimicrobial resistance genes by phages could underlie the origin of resistant bacteria found in food. We show that chicken meat carries a number of phages capable of transferring antimicrobial resistance. Of 243 phages randomly isolated from chicken meat, about a quarter (24.7%) were able to transduce resistance to one or more of the five antimicrobials tested into Escherichia coli ATCC 13706 (DSM 12242). Resistance to kanamycin was transduced the most often, followed by that to chloramphenicol, with four phages transducing tetracycline resistance and three transducing ampicillin resistance. Phages able to transduce antimicrobial resistance were isolated from 44% of the samples of chicken meat that we tested. The statistically significant (P ؍ 0.01) relationship between the presence of phages transducing kanamycin resistance and E. coli isolates resistant to this antibiotic suggests that transduction may be an important mechanism for transferring kanamycin resistance to E. coli. It appears that the transduction of resistance to certain antimicrobials, e.g., kanamycin, not only is widely distributed in E. coli isolates found on meat but also could represent a major mechanism for resistance transfer. The result is of high importance for animal and human health.
The interaction between bacteriophages, bacteria and the human host as a tripartite system has recently captured attention. The taxonomic diversity of bacteriophages, as a natural parasite of bacteria, still remains obscure in human body biomes, representing a so-called “viral dark matter.” Here, we isolated and characterized coliphages from blood, urine and tracheal aspirates samples collected at a tertiary care hospital in Austria. Phages were more often isolated from blood, followed by urine and tracheal aspirates. Phylogenetic analysis and genome comparisons allowed the identification of phages belonging to the Tunavirinae subfamily, and to the Peduovirus and Tequintavirus genera. Tunavirinae phages cluster together and are found in samples from 14 patients, suggesting their prevalence across a variety of human samples. When compared with other phage genomes, the highest similarity level was at 87.69% average nucleotide identity (ANI), which suggests that these are in fact a newly isolated phage species. Tequintavirus phages share a 95.90% with phage 3_29, challenging the ANI threshold currently accepted to differentiate phage species. The isolated phages appear to be virulent, with the exception of the Peduovirus members, which are integrative and seem to reside as prophages in bacterial genomes.
The objectives of this study were to evaluate the presence of thermophilic Campylobacter in feces of calves with and without diarrhea on dairy farms and to survey farm characteristics and management practices to define risk factors for the presence of Campylobacter. Fifty dairy farms were chosen based on the presence of calf diarrhea, and 50 farms in which calves were free from diarrhea served as a standard of comparison. In total, fecal samples were taken from 382 calves. Farm data and management practices were surveyed using a questionnaire on farm. Campylobacter were isolated from fecal samples and colonies were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis. Campylobacter spp., mainly Campylobacter jejuni (93% of isolated species), were detected on 33% of the farms and in 14.9% of the calves. Detection of Campylobacter did not differ between farms or between calves with and without diarrhea, although we found a tendency for calves suffering from diarrhea to shed Campylobacter more often. Calves may act as a reservoir of Campylobacter and may therefore lead to infections of other animals and humans. To define control strategies to reduce Campylobacter in calves, we identified on-farm risk factors. The presence of poultry on the farm, the time of cow-calf separation following birth, the use of an individual bucket for each calf, the feeding of waste milk, and the duration of individual housing were variables significantly associated with the appearance or absence of Campylobacter.
Human campylobacteriosis is primarily associated with poultry but also cattle. In this study, 55 Campylobacter jejuni strains isolated from 382 dairy calves’ feces were differentiated by multilocus sequence typing and tested for antimicrobial resistance. The most prevalent sequence type (ST) was ST883 (20.0%), followed by ST48 (14.5%), and ST50 (9.1%). In contrast to ST48 and ST50, ST883 has rarely been described in cattle previously. Furthermore, risk factor analysis was performed for the presence of the most prevalent STs in these calves. Multiple regression analysis revealed that the type of farm (organic vs. conventional) and calf housing (place, and individual vs. group) were identified as significantly (p < 0.05) associated with the presence of ST883 in calves, and ST50 was associated with calf diarrhea. Antimicrobial resistance was detected in 58.2% of the isolates. Most of the resistant isolates (81.3%) were resistant to more than one antimicrobial. Most frequently, resistance to ciprofloxacin (49.1%), followed by nalidixic acid (42.8%), and tetracycline (14.5%) was observed. The results of the present study support the hypothesis that dairy calves may serve as a potential reservoir for C. jejuni and pose a risk for transmission, including antimicrobial resistant isolates to the environment and to humans.
Campylobacter jejuni is a major cause of the Guillain-Barré syndrome (GBS) and related diseases. These autoimmune diseases are caused by antibodies cross-reacting with the peripheral (GBS) and central neural tissue (Miller Fisher syndrome-MFS, Bickerstaff's brainstem encephalitis-BBE), leading to acute polyneuropathy. Recently, specific gene loci in C. jejuni have been distinguished which are associated with the onset of GBS, despite a molecular or phenotypic clustering. In this study, we used PCR to analyse C. jejuni isolates of different origin (i.e. bovine, poultry, human) for these genes. A total of 196 isolates were tested for cst-II and neuA. Of these, 101 isolates harboured the cst-II locus and 102 the neuA locus. Eighty-six isolates (44%) hold both genes. The frequency of cst-II in different sources of isolates of bovine, poultry and human isolates did not vary significantly (52, 50 and 52%, respectively). In contrast, the neuA locus was less often found in poultry isolates. Two human strains-from a family outbreak of campylobacteriosis (in 1989 in Austria) in which one person developed MFS-harboured both genes. Thus, although only one in more than 3000 patients with Campylobacter-associated enteritis develop GBS, about half of Campylobacter jejuni strains found in different environments are possibly able to cause GBS. These strains almost equally distributed in bovine, poultry and human isolates. Our results suggest that isolates associated with GBS are not selected by environmental or host-specific factors. Accordingly, this study indicates that host factors such as humoral and cellular immunity are possibly responsible for the development of these autoimmune diseases.
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